Comparative study of the ciliary body and iris morphology in the anterior eye chamber of five different vertebrate classes.

One crucial component of the optical system is the ciliary body (CB). This body secretes the aqueous humour, which is essential to maintain the internal eye pressure as well as the clearness of the lens and cornea. The histological study was designed to provide the morphological differences of CB and iris in the anterior eye chambers of the following vertebrate classes: fish (grass carp), amphibians (Arabian toad), reptiles (semiaquatic turtle, fan-footed gecko, ocellated skink, Egyptian spiny-tailed lizard, Arabian horned viper), birds (common pigeon, common quail, common kestrel), and mammals (BALB/c mouse, rabbit, golden hamster, desert hedgehog, lesser Egyptian jerboa, Egyptian fruit bat). The results showed distinct morphological appearances of the CB and iris in each species, ranging from fish to mammals. The present comparative study concluded that the morphological structure of the CB and iris is the adaptation of species to either their lifestyle or survival in specific habitats.

Effects of silver nanoparticles prepared by aqueous extract of Ferula communis on the developing mouse embryo after maternal exposure.

Green synthesis of silver nanoparticles (AgNPs) from aqueous silver nitrate has been achieved using an extract of Ferula communis leaf as a capping, reducing, and stabilizing agent. The formation and stability of the green synthesized silver nanoparticles in the colloidal solution were monitored by absorption measurements. Silver nanoparticles were characterized by different analyses such as X-ray diffraction (XRD), energy dispersive spectroscopy (EDS), and FT-IR spectroscopy. The average particle size of silver nanoparticles was determined by high-resolution transmission electron microscopy (HRTEM) and scanning electron microscopy (SEM) analyses. In this experiment, pregnant female mice were divided into four groups (G); G1 was the control and received phosphate-buffered saline, G2 received orally aqueous extract of F. communis leaf, G3 received orally AgNPs chemically prepared by NaBH4, and G4 received orally AgNPs prepared by aqueous extract of F. communis leaf. The diameter of AgNPs was 20 nm. AgNPs exhibited good catalytic reduction ability toward methyl orange in the presence of sodium borohydride with a rate constant of 2.95 x 10-4 s-1. The results revealed the occurrence of resorbed embryos in G2, G3, and G4 with different percentages. The livers of mothers and embryos at E14.5 in G2, G3, and G4 showed different levels of histopathological alteration and increase in GFAP and CTGF expressions compared with the control group. The study concluded that the oral administration of small-sized AgNPs (20 nm) prepared by Ferula extract had less toxicity than those prepared by the chemical method.

CCN2/CTGF tip the balance of growth factors towards TGF-ß2 in primary open-angle glaucoma.

TGF-ß2 is the predominant TGF-ß isoform within the eye. One function of TGF-ß2 is to provide the eye with immune protection against intraocular inflammation. The beneficial function of TGF-ß2 within the eye must be under tight control of a network of different factors. A disbalance of the network can result in different eye diseases. In Primary Open-Angle Glaucoma (POAG), one of the leading causes of irreversible blindness worldwide, TGF-ß2 is significantly elevated in the aqueous humor and antagonistic molecules like BMPs are reduced. The changes provoke an altering of the quantity and quality of the extracellular matrix and the actin cytoskeleton in the outflow tissues, leading to an increased outflow resistance and thereby to an increased intraocular pressure (IOP), the major risk factor for primary open-angle glaucoma. The pathologic effect of TGF-ß2 in primary open-angle glaucoma is mainly meditated by CCN2/CTGF. CCN2/CTGF can modulate TGF-ß and BMP signaling by direct binding. The eye specific overexpression of CCN2/CTGF caused an increase in IOP and led to a loss of axons, the hallmark of primary open-angle glaucoma. CCN2/CTGF appears to play a critical role in the homeostatic balance of the eye, so we investigated if CCN2/CTGF can modulate BMP and TGF-ß signaling pathways in the outflow tissues. To this end, we analyzed the direct effect of CCN2/CTGF on both signaling pathways in two transgenic mouse models with a moderate (ßB1-CTGF1) and a high CCN2/CTGF (ßB1-CTGF6) overexpression and in immortalized human trabecular meshwork (HTM) cells. Additionally, we investigate whether CCN2/CTGF mediates TGF-ß effects via different pathways. We observed developmental malformations in the ciliary body in ßB1-CTGF6 caused by an inhibition of the BMP signaling pathway. In ßB1-CTGF1, we detected a dysregulation of the BMP and TGF-ß signaling pathways, with reduced BMP activity and increased TGF-ß signaling. A direct CCN2/CTGF effect on BMP and TGF-ß signaling was shown in immortalized HTM cells. Finally, CCN2/CTGF mediated its effects on TGF-ß via the RhoA/ROCK and ERK signaling in immortalized HTM cells. We conclude that CCN2/CTGF functions as a modulator of the homeostatic balance of BMP and TGF-ß signaling pathways, which is shifted in primary open-angle glaucoma.

Developmental neurotoxicity after penconazole exposure at embryo pre- and post-implantation in mice.

The present study was conducted to evaluate the effect of penconazole fungicide at a low dose (2.5 mg/kg b.w.) during embryogenesis in either the pre- or post-implantation of embryos. Females were determined pregnant according to the presence of vaginal plug and then grouped into control and penconazole-exposures at high doses (30, 20, 10, and 5 mg/kg b.w.). These high doses provoked foetoresorptionin the first experiment. Thus, a low dose (2.5 mg/kg b. w.) was used in either the pre- or post-implantation of embryos to clarify the embryotoxicity without mortality on the developing brain and eye. Results indicate a developmental delay of the cerebral hemisphere, hippocampus, cerebellum (lobulation) and induced retinopathy during eye development in post-implantation of penconazole treated group. Also, the effect of penconazole at low dose provoked a decrease in the expression of α-synuclein in the hippocampus, cerebellum, and ganglion cell layer of the developing brain and eye. In conclusion, exposure to PEN fungicide during pregnancy at a dose (2.5 mg/kg) induces alterations in the developing brain and eye tissues.

Immunohistochemical localization of alpha-synuclein in the retina of some nocturnal and diurnal animals.

Although alpha-synuclein has been reported to participate in neurodegenerative diseases, the actual normal biological function of alpha-synuclein remains unclear. I investigated the correlation of alpha-synuclein expression with nocturnal and diurnal activity for various species. Hematoxylin and eosin staining, periodic acid-Schiff's reaction (PAS) and immunohistochemistry of alpha-synuclein expression were performed for the retinas of diurnal, nocturnal, nocturnal with diurnal activity species. I found different intensity of alpha-synuclein expression in the retinal layers. I found alpha-synuclein expression in the outer segment of the photoreceptor layer in the diurnal studied species and absence of alpha-synuclein expression in the compartments of photoreceptor layer in the retina of nocturnal species. I found localization of alpha-synuclein in the inner and outer segments of photoreceptors of the retina of nocturnal with diurnal activity species. The retinas of diurnal animals exhibited glycogen in the paraboloid structure in the inner segment of the photoreceptor layer. The retinas of nocturnal and nocturnal with diurnal activity species were devoid of glycogen in the photoreceptor layer. I conclude that the function of alpha-synuclein is more related to diurnal than to nocturnal species.

Toxicological potential of penconazole on early embryogenesis of white mice Mus musculus in either pre- or post-implantation exposure.

The present investigation was conducted to evaluate the effect of penconazole (PEN) fungicide on early embryogenesis of white mice. In the first experiment, 48 pregnant females were divided into different groups; the first group is control (G1). The second group (G2) was treated daily with PEN (30-, 20-, 10-, 5-mg/kg BW). The third group (G3) was treated with PEN (5-mg/kg BW; day after the other day). The fourth group (G4) was treated with PEN (2.5-mg/kg BW daily) during pre-implantation stage (from the 1st to the 4th day of gestation). The fifth group (G5) was treated with PEN (2.5-mg/kg BW daily) during post-implantation (from the 5th to the 8th day of gestation). The pregnant females were sacrificed at the 14th day of gestation. In the second experiment, 63 pregnant females were classified into control, PEN-treated during pre-implantation period (2.5-mg/kg BW), and PEN-administered during post-implantation period (2.5-mg/kg BW). Each group was sacrificed at stages E6.5, E7.5, E8.5, E9.5, E11.5, E14.5, and E18.5. The high doses of PEN in the first experiment showed failed pregnancy, foetoresorption, and embryo disorganization. High doses of PEN induce alterations in the uterus tissue at the level of histology and immunohistochemistry for the expression of TGFß2, TNFR2, Caspase 10, and HSP70. The low doses of PEN in the second experiment showed upregulated expression of TGFß2, TNFR2, Caspase 10, and HSP70 at stages E6.5 and E7.5. In conclusion, PEN was found to alter the suitable uterine environment for proper implantation and development at the levels of histological and immunohistochemical that could create a risk during the full course of embryogenesis.

Teratogenicity and neurotoxicity effects induced by methomyl insecticide on the developmental stages of Bufo arabicus.

Methomyl (MET) is a carbamate insecticide that has been widely used to protect the crop against insects as an alternative for organophosphorus insecticide. Thus the present study aims to evaluate the potential toxic effects of MET on the developmental stages of Bufo arabicus. Tadpoles were classified into three stages (25, 37, 40). Every stage was divided into two groups, control and MET-treated group (10 ppm for two weeks) after LC50 determination in acute toxicity test for 96 h. Control and MET-treated larvae were examined at the level of morphological, histological, skeleton deformities and immunohistochemical labeling of alpha-synuclein in the spinal cord and dorsal root ganglion. MET-exposed larvae showed hyperactivity, extreme agitation, abnormal swimming and kinking tail as compared to control. Alizarin Red S-Alcian blue staining showed scoliosis in MET-treated tadpoles at 25 and 37 stages; kyphosis, retarded tail regression and reduced ossification of the phalanges of digits for both fore-and hind limbs were noted in MET-exposed tadpoles at 40 stage as compared to control. Histopathological changes in myotomes, notochord and spinal cord were shown in MET-exposed tadpoles as compared to control. Immunohistochemical examination showed an over expression of alpha-synuclein either in the neurons of the spinal cord or in the dorsal root ganglion of MET-exposed tadpoles at stage 40 as compared to control. The present study concluded that MET insecticide induces malformation and teratogenicity effects which were accompanied by neurodegenerative effects for the neurons either in the spinal cord or in the dorsal root ganglion.

Synthesis and effect of a new Terbium gibberellic complex on the histopathological alteration induced by Gibberellic acid on liver and kidney of mice Mus musculus.

The objective of this study was to synthesize Gibberellic lanthanide complex and evaluate its biological activity to reduce the Gibberellic acid toxicity on liver and kidney. The new bis(Gibberellic)-nitro-terbium(III) complex was characterized by different analytical methods: elemental analyses, UV-Vis, molar ratio, fluorescence, FT-IR, and TGA-DTA measurements. Thirty newborns were classified into three groups control, Gibberellic acid, and Terbium gibberellic acid complex. Livers and kidneys of studied groups proceed for general histology and immunohistochemical staining of Cyr61, cytochrome C, and TNFR2. From the absorption titration measurements, the binding constants of DNA with Tb(III)-(GA)2 complex and free ligand were found to be 3.9 × 104 and 2.1 × 103  m-1 , respectively, with the stoichiometry of 1:1. Hypochromism was observed from the absorption titration experiment which indicates the intercalation of Tb(III)-(GA)2 complex between the base pairs of DNA. Gibberellic acid-treated group showed alteration in the histological picture of livers and kidneys that accompanied with the reduction in the expression of Cyr61, cyt C, and TNFR2. The amelioration was observed in Gibberellic acid complex with Terbium group. The study concluded that Terbium gibberellic complex is less dangerous effects than Gibberellic acid alone.

Expression of alpha-synuclein during eye development of mice (Mus musculus), chick (Gallus gallus domisticus) and fish (Ctenopharyngodon idella) in a comparison study.

Synucleins are small proteins associated with neurodegenerative diseases, alpha-synuclein is a Parkinson's disease-linked protein of ubiquitous expression in the central nervous system. This study aimed at the localization of alpha-synuclein during eye development of mice (Mus musculus), chick (Gallus gallus domisticus) and fish (Ctenopharyngodon idella) by immunohistochemical staining in a comparison study. The results showed that alpha-synuclein expression increased gradually with the development of ciliary body, iris, retina and cornea of mice at E17, P1, P3, P7 and chick at E5, E10, E15 with unequal appearance of alpha-synuclein expression. Also, it was not detected in iridocorneal angle during eye development of mice and chick. Alpha-synuclein expression during fish eye development at P10, P15, P20 was not detected either in the ciliray body or Iris regions and it was pronounced with sharp signals in the highly specialized tissue of the iridocorneal angle at P20. Also, the expression was gradually increased from P15 to P20 in fish retina and cornea. The pattern of expression and distribution of alpha-synuclein during the development of ciliary body and iris of mice, chick and fish has not been previously characterized, The data concluded that alpha-synuclein has important cellular function during eye development of studied animals.

Connective tissue growth factor causes glaucoma by modifying the actin cytoskeleton of the trabecular meshwork.

The most critical risk factor for optic nerve damage in cases of primary open-angle glaucoma (POAG) is an increased intraocular pressure (IOP) caused by a resistance to aqueous humor outflow in the trabecular meshwork (TM). The molecular pathogenesis of this increase in outflow resistance in POAG has not yet been identified, but it may involve transforming growth factor TGF-ß2, which is found in higher amounts in the aqueous humor of patients with POAG. Connective tissue growth factor (CTGF) is a TGF-ß2 target gene with high constitutive TM expression. In this study, we show that either adenoviral-mediated or transgenic CTGF overexpression in the mouse eye increases IOP and leads to optic nerve damage. CTGF induces TM fibronectin and α-SMA in animals, whereas actin stress fibers and contractility are both induced in cultured TM cells. Depletion of CTGF by RNA interference leads to a marked attenuation of the actin cytoskeleton. Rho kinase inhibitors cause a reversible decline in the IOP of CTGF-overexpressing mice to levels seen in control littermates. Overall, the effects of CTGF on IOP appear to be caused by a modification of the TM actin cytoskeleton. CTGF-overexpressing mice provide a model that mimics the essential functional and structural aspects of POAG and offer a molecular mechanism to explain the increase of its most critical risk factor.